TY - JOUR

T1 - Dermal fibroblasts have different extracellular matrix profiles induced by TGF-beta, PDGF and IL-6 in a model for skin fibrosis

AU - Juhl, Pernille

AU - Bondesen, Sandie

AU - Hawkins, Clare Louise

AU - Karsdal, Morten Asser

AU - Bay-Jensen, Anne-Christine

AU - Davies, Michael Jonathan

AU - Siebuhr, Anne Sofie

PY - 2020

Y1 - 2020

N2 - Different stimulants might induce different extracellular matrix profiles. It is essential to gain an understanding and quantification of these changes to allow for focused anti-fibrotic drug development. This study investigated the expression of extracellular matrix by dermal fibroblast mimicking fibrotic skin diseases as SSc using clinically validated biomarkers. Primary healthy human dermal fibroblasts were grown in media containing FICOLL. The cells were stimulated with PDGF-AB, TGF-beta 1, or IL-6. Anti-fibrotic compounds (iALK-5, Nintedanib) were added together with growth factors. Biomarkers of collagen formation and degradation together with fibronectin were evaluated by ELISAs in the collected supernatant. Immunohistochemical staining was performed to visualize fibroblasts and proteins, while selected gene expression levels were examined through qPCR. TGF-beta and PDGF, and to a lesser extent IL-6, increased the metabolic activity of the fibroblasts. TGF-beta primarily increased type I collagen and fibronectin protein and gene expression together with alpha SMA. PDGF stimulation resulted in increased type III and VI collagen formation and gene expression. IL-6 decreased fibronectin levels. iALK5 could inhibit TGF-beta induced fibrosis while nintedanib could halt fibrosis induced by TGF-beta or PDGF. Tocilizumab could not inhibit fibrosis induced in this model. The extent and nature of fibrosis are dependent on the stimulant. The model has potential as a pre-clinical model as the fibroblasts fibrotic phenotype could be reversed by an ALK5 inhibitor and Nintedanib.

AB - Different stimulants might induce different extracellular matrix profiles. It is essential to gain an understanding and quantification of these changes to allow for focused anti-fibrotic drug development. This study investigated the expression of extracellular matrix by dermal fibroblast mimicking fibrotic skin diseases as SSc using clinically validated biomarkers. Primary healthy human dermal fibroblasts were grown in media containing FICOLL. The cells were stimulated with PDGF-AB, TGF-beta 1, or IL-6. Anti-fibrotic compounds (iALK-5, Nintedanib) were added together with growth factors. Biomarkers of collagen formation and degradation together with fibronectin were evaluated by ELISAs in the collected supernatant. Immunohistochemical staining was performed to visualize fibroblasts and proteins, while selected gene expression levels were examined through qPCR. TGF-beta and PDGF, and to a lesser extent IL-6, increased the metabolic activity of the fibroblasts. TGF-beta primarily increased type I collagen and fibronectin protein and gene expression together with alpha SMA. PDGF stimulation resulted in increased type III and VI collagen formation and gene expression. IL-6 decreased fibronectin levels. iALK5 could inhibit TGF-beta induced fibrosis while nintedanib could halt fibrosis induced by TGF-beta or PDGF. Tocilizumab could not inhibit fibrosis induced in this model. The extent and nature of fibrosis are dependent on the stimulant. The model has potential as a pre-clinical model as the fibroblasts fibrotic phenotype could be reversed by an ALK5 inhibitor and Nintedanib.

KW - GROWTH-FACTOR-BETA

KW - SYSTEMIC-SCLEROSIS

KW - I COLLAGEN

KW - INTERLEUKIN-6

KW - ACTIVATION

KW - LIVER

KW - VITRO

U2 - 10.1038/s41598-020-74179-6

DO - 10.1038/s41598-020-74179-6

M3 - Journal article

C2 - 33057073

VL - 10

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

M1 - 17300

ER -